alpha-Crystallin and small heat-shock proteins (sHSP) show sequence similarity along a stretch of 100 amino acids referred to as the alpha-crystallin domain. The presence of this domain is associated with the ability of sHSP to assemble into oligomeric structures and with their function as molecular chaperones. The purpose of this work is to determine whether highly conserved sequences in the alpha-crystallin domain of human heat shock protein 27 (HSP 27) and alphaA-crystallin are involved in contacts between subunits in the oligomeric structure. Sequential single cysteine mutants were constructed along the sequence between residues 109 and 120 in alpha-crystallin and along the corresponding sequence in HSP 27. The mutants were purified, spin-labeled and analyzed by Electron Paramagnetic Resonance (EPR) spectroscopy. Analysis of the sequence-specific environment of the attached spin labels reveals a periodicity of two consistent with the presence of a beta-strand along the sequences. The EPR spectral line shapes indicate that in both proteins, spin labels on different monomers are brought into close proximity in the oligomeric structure suggesting that equivalent strands from different subunits are interacting along an interface. A similar pattern of interactions is obtained in a mixed oligomer of alphaA-crystallin and HSP 27. Our data indicate a similar quarternary structure for alphaA-crystalline and HSP 27. Because the studied sequence is highly conserved across the sHSP family, we propose that the oligomeric structure of these proteins involves a conserved two-fold isologous association.?

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Biotechnology Resource Grants (P41)
Project #
5P41RR001008-23
Application #
6279888
Study Section
Project Start
1998-04-01
Project End
1999-02-28
Budget Start
1997-10-01
Budget End
1998-09-30
Support Year
23
Fiscal Year
1998
Total Cost
Indirect Cost
Name
Medical College of Wisconsin
Department
Type
DUNS #
073134603
City
Milwaukee
State
WI
Country
United States
Zip Code
53226
Mao, Li; Liu, Yu-Xiang; Huang, Chun-Hua et al. (2015) Intrinsic Chemiluminescence Generation during Advanced Oxidation of Persistent Halogenated Aromatic Carcinogens. Environ Sci Technol 49:7940-7
Shan, Guo-Qiang; Yu, Ao; Zhao, Chuan-Fang et al. (2015) A combined experimental and computational investigation on the unusual molecular mechanism of the Lossen rearrangement reaction activated by carcinogenic halogenated quinones. J Org Chem 80:180-9
Li, Yan; Huang, Chun-Hua; Liu, Yu-Xiang et al. (2014) Detoxifying polyhalogenated catechols through a copper-chelating agent by forming stable and redox-inactive hydrogen-bonded complexes with an unusual perpendicular structure. Chemistry 20:13028-33
Sheng, Zhi-Guo; Huang, Wei; Liu, Yu-Xiang et al. (2013) Ofloxacin induces apoptosis via ?1 integrin-EGFR-Rac1-Nox2 pathway in microencapsulated chondrocytes. Toxicol Appl Pharmacol 267:74-87
Sheng, Zhi-Guo; Huang, Wei; Liu, Yu-Xiang et al. (2013) Bisphenol A at a low concentration boosts mouse spermatogonial cell proliferation by inducing the G protein-coupled receptor 30 expression. Toxicol Appl Pharmacol 267:88-94
Shao, Jie; Huang, Chun-Hua; Kalyanaraman, Balaraman et al. (2013) Potent methyl oxidation of 5-methyl-2'-deoxycytidine by halogenated quinoid carcinogens and hydrogen peroxide via a metal-independent mechanism. Free Radic Biol Med 60:177-82
Sheng, Zhi-Guo; Li, Yan; Fan, Rui-Mei et al. (2013) Lethal synergism between organic and inorganic wood preservatives via formation of an unusual lipophilic ternary complex. Toxicol Appl Pharmacol 266:335-44
Qin, Hao; Huang, Chun-Hua; Mao, Li et al. (2013) Molecular mechanism of metal-independent decomposition of lipid hydroperoxide 13-HPODE by halogenated quinoid carcinogens. Free Radic Biol Med 63:459-66
Huang, Chun-Hua; Shan, Guo-Qiang; Mao, Li et al. (2013) The first purification and unequivocal characterization of the radical form of the carbon-centered quinone ketoxy radical adduct. Chem Commun (Camb) 49:6436-8
Liddle, Brendan J; Wanniarachchi, Sarath; Hewage, Jeewantha S et al. (2012) Electronic communication across diamagnetic metal bridges: a homoleptic gallium(III) complex of a redox-active diarylamido-based ligand and its oxidized derivatives. Inorg Chem 51:12720-8

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